Molecular analysis of gene abnormalities in myasthenia gravis.

重症肌无力基因异常的分子分析。

基本信息

批准号：
02454224
负责人：
ARIMORI Shigeru
金额：
$ 4.48万
依托单位：
Tokai University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

1. Genomic DNAs of Japanese healthy individuals were digested with endonuclease Kpn I, Pst I, Pvu II, Hint I, and Sac I. Southern blot analysis of these DNA digests was performed by using calf acetylcholine delta/gamma subunit CDNA as a probe.DNA polymorphysm was found only in Pvu II-digested DNAs (1.7 kb and 1.9 kb bands) in Japanese. The frequency of 1.7 kb/1.7 kb was 85% (83/98) and of 1.7 kb/1.9 kb was 15% (15/98). The frequencies of these bands in Japanese myasthenia gravis patients were 88% and 12%, respectively. Statistically significant association was not demonstrated between myasthenia gravis and RFLPs of acetylcholine receptor delta/gamma subunit gene.2. BUF/Mna rats develop spontaneous benign thymomas of epithelial origin with nearly 100% incidence. We purified satellite I DNA from BUF/Mna rat thymoma and constructed recombinant plasmid library. We screened 1200 clones by using satellite I DNA probe and isolated 4 unusual clones. Nucleotide sequence analysis of these clones revealed many deletions, insertions and base substitutions, and these mutations created unusual repeat units. The unusual structures of satellite I DNA subunit were created by change of Hint I sequence to Eco RI sequence in monomeric and dimeric-subunit. The nucleotide sequence TGGGAAC, which is strictly conserved in normal repeat unit was completely lost in all these clones. Control experiment was done in ACI rat DNA. Out of 1200 clones examined, no unusual satellite I DNA sequence was found. The generation of abnormal satellite I DNA may provide the molecular basis of chromosome instability in thymoma cells of BUF/Mna rats.

1。日本健康个体的基因组DNA用核酸内切酶KPN I，PST I，PVU II，提示I和SACI。对这些DNA消化的Southern印迹分析是通过使用小牛乙酰胆碱delta/gamma subunit cdna作为Probe.dna Polymorphymphysm and polymorphysm and p kb kb kb kb kb kb kb kb kb kb kb kb kb kb kb ii（乐队）日语。 1.7 kb/1.7 kb的频率为85％（83/98）和1.7 kb/1.9 kb的频率为15％（15/98）。这些乐队在日本肌无力的肌无力患者中的频率分别为88％和12％。在肌动脉症和乙酰胆碱受体Delta/Gamma亚基基因的RFLP之间没有证明具有统计学意义的关联。2。 BUF/MNA大鼠出现上皮起源的自发良性胸腺瘤，发病率将近100％。我们从BUF/MNA大鼠胸腺瘤和构建的重组质粒文库中纯化了卫星I DNA。我们使用卫星I DNA探针筛选了1200个克隆，并隔离了4个不寻常的克隆。这些克隆的核苷酸序列分析揭示了许多缺失，插入和基本取代，这些突变产生了异常的重复单位。卫星I DNA亚基的异常结构是通过将i序列变为单体和二聚体 - 亚基中的ECO RI序列而产生的。在所有这些克隆中，在正常重复单元中严格保守的核苷酸序列TGGGAAC完全丢失。对照实验是在ACI大鼠DNA中进行的。在检查的1200个克隆中，没有发现异常的卫星I DNA序列。异常卫星I DNA的产生可能提供BUF/MNA大鼠胸腺瘤细胞中染色体不稳定性的分子基础。